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Supplementary Figure Legends
Supplementary Figure S1 Virus accumulation in N. benthamiana plants treated with various concentrations of
BL. The fifth or sixth (local) leaf were treated with different concentrations of BL, 12 h later, the seventh or
eighth (upper) leaf was inoculated with TMV-GFP. (a) Images of GFP fluorescence in upper leaves. Pictures
were taken under a UV light at 1, 3 and 5 dpi. (b) qRT-PCR analysis of TMV mRNA accumulation levels in
inoculated leaves collected at 1, 3 and 5 dpi. Bars represent mean and standard deviation of values obtained from
three biological replicates per genotype and time point. Significant differences (P< 0.05) are denoted by different
lowercase letters.
Supplementary Figure S2 BRs inhibit viral replication and spread in the non-inoculated upper leaves. (a)
Images of GFP fluorescence in non-inoculated upper leaves. Pictures were taken under a UV light at 12 dpi. (b)
qRT-PCR analysis of TMV mRNA accumulation levels in non-inoculated leaves at 12 dpi. Bars represent mean
and standard deviation of values obtained from three biological replicates per genotype and time point.
Significant differences (P< 0.05) are denoted by different lowercase letters.
Supplementary Figure S3 Relative expression of NbCPD and NbDWARF in local and upper leaves after 12 h of
local BL treatment. Bars represent mean and standard deviation of values obtained from three biological
replicates per genotype and time point. Significant differences (P< 0.05) are denoted by different lowercase
letters.
Supplementary Figure S4 Time course of H2O2 and NO contents in local or upper leaves after local leaves were
treated with water or BL. Bars represent mean and standard deviation of values obtained from three biological
replicates per genotype and time point.
Supplementary Figure S5 Average GFP fluorescence intensity in Figure 2a. Bars represent mean and standard
deviation of values obtained from three biological replicates per genotype and time point. Significant differences
(P< 0.05) are denoted by different lowercase letters.
Supplementary Figure S6 Pharmacologic analysis of NADPH oxidase in BR-induced H2O2 production. (a)
H2O2 levels in local and upper leaves stained with H2DCF-DA were detected by a fluorescence microscope.
Local or upper leaves were pretreated with DPI for 4 h, then local leaves were treated with BL for another 12 h.
(b) Quantitative measurements of H2O2 contents in leaves shown in (a). Bars represent mean and standard
deviation of values obtained from three biological replicates per genotype and time point. Significant differences
(P< 0.05) are denoted by different lowercase letters. L, local leaves; U, upper leaves.
Supplementary Figure S7 Confirmation of the NbRBOHA, NbRBOHB, NbNOA1, NbNR and NbBRI1 genes
silencing in N. benthamiana plants. qRT-PCR analysis was conducted using total RNA extracted from plants
inoculated with Agrobacterium GV2260 carrying TRV-target genes and the corresponding non-silenced TRV:00
infected plants. Bars represent mean and standard deviation of values obtained from three biological repeats.
Supplementary Figure S8 Average GFP fluorescence intensity in Figure 4a. Bars represent mean and standard
deviation of values obtained from three biological replicates per genotype and time point. Significant differences
(P< 0.05) are denoted by different lowercase letters.
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Supplementary Figure S9 Pharmacologic analysis of NOS and NR in BR-induced NO production. (a) NO
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Supplementary Table S1 Primers used for construction of VIGS and RNAi vectors.
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Supplementary Table S2 Primers used for quantitative real-time PCR analysis of gene expression.
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levels in local and upper leaves stained with DAF-FM-DA were detected by a fluorescence microscope. Local or
upper leaves were pretreated with L-NAME or tungstate for 4 h, then local leaves were treated with BL for
another 12 h. (b) Quantitative measurements of NO contents in leaves shown in (a). Bars represent mean and
standard deviation of values obtained from three biological replicates per genotype and time point. Significant
differences (P< 0.05) are denoted by different lowercase letters. L, local leaves; U, upper leaves.
Supplementary Figure S10 Confirmation of NbRBOHB and NbNR transient silencing efficiency. (a) Schematic
illustration of intron-containing hairpin RNA recombinant vector carrying target genes fragment. (b) qRT-PCR
analysis of NbRBOHB and NbNR expression in leaves infiltrated with empty or RNAi vectors. Bars represent
mean and standard deviation of values obtained from three biological repeats.
Supplementary Figure S11 Virus accumulation in NbRBOHB-silenced plants in the presence of various
concentrations of SNP. (a) Images of GFP fluorescence in upper leaves of NbRBOHB-silenced plants. Local
leaves were treated with BL, at the same time, upper leaves were spraied with different concentrations of SNP.
12 h later, upper leaves were inoculated with TMV-GFP. Images of virus accumulation were taken at 3 dpi. (b)
qRT-PCR analysis of TMV mRNA accumulation levels in inoculated leaves collected at 3 dpi. Bars represent
mean and standard deviation of values obtained from three biological replicates per genotype and time point.
Significant differences (P< 0.05) are denoted by different lowercase letters.
Supplementary Figure S12 H2O2 and SNP can rescue BR-induced systemic virus resistance in
NbBRI1-silenced plants. (a) Images of GFP fluorescence in upper leaves of NbBRI1-silenced and control plants.
Local leaves were treated with BL, at the same time, upper leaves were treated with H2O2 or SNP. 12 h later,
upper leaves were inoculated with TMV-GFP. Images of virus accumulation were taken at 3 dpi. (b) qRT-PCR
analysis of TMV mRNA accumulation levels in inoculated leaves collected at 3 dpi. Bars represent mean and
standard deviation of values obtained from three biological replicates per genotype and time point. Significant
differences (P< 0.05) are denoted by different lowercase letters.
Supplementary Figure S13 Relative expression of NbRBOHA, NbRBOHB, NbNOA1 and NbNR in local and
upper leaves after 12 h of local BL treatment. Bars represent mean and standard deviation of values obtained
from three biological replicates per genotype and time point. Significant differences (P< 0.05) are denoted by
different lowercase letters